Alternative buffer systems in biopharmaceutical formulations and their effect on protein stability

Lebar, Blaž (author), Zidar, Mitja (author), Mravljak, Janez (author), Šink, Roman (author), Žula, Aleš (author), Pajk, Stane (author)

The formulation of biopharmaceutical drugs is designed to eliminate chemical instabilities, increase conformational and colloidal stability of proteins, and optimize interfacial stability. Among the various excipients involved, buffer composition plays a pivotal role. However, conventional buffers like histidine and phosphate buffers may not always be the optimal choice for all monoclonal antibodies (mAbs). In this study, we investigated the effects of several alternative buffer systems on seven different mAbs, exploring various combinations of ionic strengths, concentrations of the main buffer component, mAb concentrations and stress conditions. Protein stability was assessed by analysing soluble aggregate formation through size exclusion chromatography. At low protein concentration, protein instability after temperature stress was exclusively observed in the bis-TRIS/glucuronate buffer. Conversely, freeze-thaw stress led to significant increase in aggregate formation in tested formulations, highlighting the efficacy of several alternative buffers, particularly arginine/citrate, in preserving protein stability. Under temperature stress, the introduction of arginine to histidine buffer systems provided additional stabilization, while the addition of lysine resulted in protein destabilization. Similarly, the incorporation of arginine into histidine/HCl buffer further enhanced protein stability during freeze thaw cycles. At high protein concentration, the histidine/citrate buffer emerged as one of the most optimal choices for addressing temperature and light induced stress. The efficacy of histidine buffers in combating light stress might be attributed to the light absorbing properties of histidine molecules. Our findings demonstrate that the development of biopharmaceutical formulations should not be confined to conventional buffer systems, as numerous alternative options exhibit comparable or even superior performance.

• Language: English
• Keywords: biopharmaceuticals, buffer, stress, aggregates, alternative buffers, stability
• Work type: Article
• Typology: 1.01 - Original Scientific Article
• Organization: FFA - Faculty of Pharmacy
• Publication status: Published
• Publication version: Version of Record
• Year: 2024
• Number of pages: Str. 479–493
• Numbering: Vol. 74, no. 3
• PID: 20.500.12556/RUL-163471
• UDC: 615.4:54
• ISSN on article: 1846-9558
• DOI: 10.2478/acph-2024-0022
• COBISS.SI-ID: 197188099
• Publication date in RUL: 07.10.2024

Record is a part of a journal

• Title: Acta pharmaceutica
• Shortened title: Acta pharm.
• Publisher: Croatian Pharmaceutical Society
• ISSN: 1846-9558
• COBISS.SI-ID: 3817585

Licences

• License: CC BY-NC 4.0, Creative Commons Attribution-NonCommercial 4.0 International
• Link: http://creativecommons.org/licenses/by-nc/4.0/
• Description: A creative commons license that bans commercial use, but the users don’t have to license their derivative works on the same terms.

Secondary language

• Language: Slovenian
• Keywords: biofarmacevtika, pufer, stres, agregati, alternativni pufri, stabilnost, farmacevtska kemija

Projects

• Funder: Other - Other funder or multiple funders
• Funding programme: Novartis Pharmaceutical Manufacturing LLC

• Funder: ARIS - Slovenian Research and Innovation Agency
• Project number: P1-0208
• Name: Farmacevtska kemija: načrtovanje, sinteza in vrednotenje učinkovin